A Proteomic Investigation to Discover Candidate Proteins Involved in Novel Mechanisms of 5-Fluorouracil Resistance in Colorectal Cancer

One of the main obstacles to therapeutic success in colorectal cancer (CRC) is the development of acquired resistance to treatment with drugs such as 5-fluorouracil (5-FU). Whilst some resistance mechanisms are well known, it is clear from the stasis in therapy success rate that much is still unknow...

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Published inCells (Basel, Switzerland) Vol. 13; no. 4; p. 342
Main Authors Ortega Duran, Mario, Shaheed, Sadr ul, Sutton, Christopher W., Shnyder, Steven D.
Format Journal Article
LanguageEnglish
Published Switzerland MDPI AG 01.02.2024
Subjects
5-Fluorouracil
Amino acids
Cancer therapies
Causes of
CD44 antigen
CD63 antigen
Cell culture
Cell cycle
Cell Line, Tumor
Colorectal cancer
Colorectal carcinoma
Colorectal Neoplasms - drug therapy
Colorectal Neoplasms - metabolism
Dosage and administration
Drug development
Drug resistance
drug resistance mechanisms
Drug Resistance, Neoplasm
Drug therapy
Fluorouracil
Fluorouracil - pharmacology
Fluorouracil - therapeutic use
Genomics
Health aspects
Humans
in vitro models
Labeling
Liquid chromatography
Mass spectroscopy
Metabolism
Metabolites
Mucoproteins
Oncogene Proteins
Peptides
Proteins
Proteomics
stable isotope labelling with amino acids in cell culture (SILAC)
Tumorigenesis
United Kingdom
United Kingdom > UK
5-fluorouracil
colorectal cancer
drug resistance mechanisms
proteomics
in vitro models
stable isotope labelling with amino acids in cell culture (SILAC)
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Abstract One of the main obstacles to therapeutic success in colorectal cancer (CRC) is the development of acquired resistance to treatment with drugs such as 5-fluorouracil (5-FU). Whilst some resistance mechanisms are well known, it is clear from the stasis in therapy success rate that much is still unknown. Here, a proteomics approach is taken towards identification of candidate proteins using 5-FU-resistant sublines of human CRC cell lines generated in house. Using a multiplexed stable isotope labelling with amino acids in cell culture (SILAC) strategy, 5-FU-resistant and equivalently passaged sensitive cell lines were compared to parent cell lines by growing in Heavy medium with 2D liquid chromatography and Orbitrap Fusion™ Tribrid™ Mass Spectrometry analysis. Among 3003 commonly quantified proteins, six (CD44, APP, NAGLU, CORO7, AGR2, PLSCR1) were found up-regulated, and six (VPS45, RBMS2, RIOK1, RAP1GDS1, POLR3D, CD55) down-regulated. A total of 11 of the 12 proteins have a known association with drug resistance mechanisms or role in CRC oncogenesis. Validation through immunodetection techniques confirmed high expression of CD44 and CD63, two known drug resistance mediators with elevated proteomics expression results. The information revealed by the sensitivity of this method warrants it as an important tool for elaborating the complexity of acquired drug resistance in CRC.
AbstractList One of the main obstacles to therapeutic success in colorectal cancer (CRC) is the development of acquired resistance to treatment with drugs such as 5-fluorouracil (5-FU). Whilst some resistance mechanisms are well known, it is clear from the stasis in therapy success rate that much is still unknown. Here, a proteomics approach is taken towards identification of candidate proteins using 5-FU-resistant sublines of human CRC cell lines generated in house. Using a multiplexed stable isotope labelling with amino acids in cell culture (SILAC) strategy, 5-FU-resistant and equivalently passaged sensitive cell lines were compared to parent cell lines by growing in Heavy medium with 2D liquid chromatography and Orbitrap Fusion™ Tribrid™ Mass Spectrometry analysis. Among 3003 commonly quantified proteins, six (CD44, APP, NAGLU, CORO7, AGR2, PLSCR1) were found up-regulated, and six (VPS45, RBMS2, RIOK1, RAP1GDS1, POLR3D, CD55) down-regulated. A total of 11 of the 12 proteins have a known association with drug resistance mechanisms or role in CRC oncogenesis. Validation through immunodetection techniques confirmed high expression of CD44 and CD63, two known drug resistance mediators with elevated proteomics expression results. The information revealed by the sensitivity of this method warrants it as an important tool for elaborating the complexity of acquired drug resistance in CRC.
One of the main obstacles to therapeutic success in colorectal cancer (CRC) is the development of acquired resistance to treatment with drugs such as 5-fluorouracil (5-FU). Whilst some resistance mechanisms are well known, it is clear from the stasis in therapy success rate that much is still unknown. Here, a proteomics approach is taken towards identification of candidate proteins using 5-FU-resistant sublines of human CRC cell lines generated in house. Using a multiplexed stable isotope labelling with amino acids in cell culture (SILAC) strategy, 5-FU-resistant and equivalently passaged sensitive cell lines were compared to parent cell lines by growing in Heavy medium with 2D liquid chromatography and Orbitrap Fusion™ Tribrid™ Mass Spectrometry analysis. Among 3003 commonly quantified proteins, six (CD44, APP, NAGLU, CORO7, AGR2, PLSCR1) were found up-regulated, and six (VPS45, RBMS2, RIOK1, RAP1GDS1, POLR3D, CD55) down-regulated. A total of 11 of the 12 proteins have a known association with drug resistance mechanisms or role in CRC oncogenesis. Validation through immunodetection techniques confirmed high expression of CD44 and CD63, two known drug resistance mediators with elevated proteomics expression results. The information revealed by the sensitivity of this method warrants it as an important tool for elaborating the complexity of acquired drug resistance in CRC.One of the main obstacles to therapeutic success in colorectal cancer (CRC) is the development of acquired resistance to treatment with drugs such as 5-fluorouracil (5-FU). Whilst some resistance mechanisms are well known, it is clear from the stasis in therapy success rate that much is still unknown. Here, a proteomics approach is taken towards identification of candidate proteins using 5-FU-resistant sublines of human CRC cell lines generated in house. Using a multiplexed stable isotope labelling with amino acids in cell culture (SILAC) strategy, 5-FU-resistant and equivalently passaged sensitive cell lines were compared to parent cell lines by growing in Heavy medium with 2D liquid chromatography and Orbitrap Fusion™ Tribrid™ Mass Spectrometry analysis. Among 3003 commonly quantified proteins, six (CD44, APP, NAGLU, CORO7, AGR2, PLSCR1) were found up-regulated, and six (VPS45, RBMS2, RIOK1, RAP1GDS1, POLR3D, CD55) down-regulated. A total of 11 of the 12 proteins have a known association with drug resistance mechanisms or role in CRC oncogenesis. Validation through immunodetection techniques confirmed high expression of CD44 and CD63, two known drug resistance mediators with elevated proteomics expression results. The information revealed by the sensitivity of this method warrants it as an important tool for elaborating the complexity of acquired drug resistance in CRC.
Audience Academic
Author Shaheed, Sadr ul
Shnyder, Steven D.
Ortega Duran, Mario
Sutton, Christopher W.
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CitedBy_id crossref_primary_10_1016_j_biopha_2024_117576
crossref_primary_10_1021_acsptsci_4c00008
crossref_primary_10_1016_j_cytogfr_2024_08_006
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Keywords 5-fluorouracil
colorectal cancer
drug resistance mechanisms
proteomics
in vitro models
stable isotope labelling with amino acids in cell culture (SILAC)
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Snippet One of the main obstacles to therapeutic success in colorectal cancer (CRC) is the development of acquired resistance to treatment with drugs such as...
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SubjectTerms 5-Fluorouracil
Amino acids
Cancer therapies
Causes of
CD44 antigen
CD63 antigen
Cell culture
Cell cycle
Cell Line, Tumor
Colorectal cancer
Colorectal carcinoma
Colorectal Neoplasms - drug therapy
Colorectal Neoplasms - metabolism
Dosage and administration
Drug development
Drug resistance
drug resistance mechanisms
Drug Resistance, Neoplasm
Drug therapy
Fluorouracil
Fluorouracil - pharmacology
Fluorouracil - therapeutic use
Genomics
Health aspects
Humans
in vitro models
Labeling
Liquid chromatography
Mass spectroscopy
Metabolism
Metabolites
Mucoproteins
Oncogene Proteins
Peptides
Proteins
Proteomics
stable isotope labelling with amino acids in cell culture (SILAC)
Tumorigenesis
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Title A Proteomic Investigation to Discover Candidate Proteins Involved in Novel Mechanisms of 5-Fluorouracil Resistance in Colorectal Cancer
URI https://www.ncbi.nlm.nih.gov/pubmed/38391955
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Volume 13
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